Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells.

Chandrasekaran A; Avci HX; Ochalek A; Rösingh LN; Molnár K; László L; Bellák T; Téglási A; Pesti K; Mike A; Phanthong P; Bíró O; Hall V; Kitiyanant N; Krause KH; Kobolák J; Dinnyés A

doi:10.1016/j.scr.2017.10.010

Back

Journal article

Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells.

Chandrasekaran A BioTalentum Ltd, Gödöllő, Hungary; Molecular Animal Biotechnology Lab, Szent István University, Gödöllő, Hungary.
Avci HX BioTalentum Ltd, Gödöllő, Hungary; Department of Anatomy, Embryology and Histology, Faculty of Medicine, University of Szeged, Szeged, Hungary.
Ochalek A BioTalentum Ltd, Gödöllő, Hungary; Molecular Animal Biotechnology Lab, Szent István University, Gödöllő, Hungary.
Rösingh LN Department of Pathology and Immunology, University of Geneva Medical School, Geneva, Switzerland.
Molnár K Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary.
László L Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary.
Bellák T BioTalentum Ltd, Gödöllő, Hungary; Department of Anatomy, Embryology and Histology, Faculty of Medicine, University of Szeged, Szeged, Hungary.
Téglási A BioTalentum Ltd, Gödöllő, Hungary.
Pesti K Opto-Neuropharmacology Group, MTA-ELTE NAP B, Budapest, Hungary; János Szentágothai Doctoral School of Neurosciences, Semmelweis University, Budapest, Hungary.
Mike A Opto-Neuropharmacology Group, MTA-ELTE NAP B, Budapest, Hungary.
Phanthong P BioTalentum Ltd, Gödöllő, Hungary; Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom Bangkok, Thailand.
Bíró O First Department of Obstetrics and Gynaecology, Semmelweis University, Budapest, Hungary.
Hall V Department of Veterinary and Animal Science, University of Copenhagen, Denmark.
Kitiyanant N Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom Bangkok, Thailand.
Krause KH Department of Pathology and Immunology, University of Geneva Medical School, Geneva, Switzerland.
Kobolák J BioTalentum Ltd, Gödöllő, Hungary.
Dinnyés A BioTalentum Ltd, Gödöllő, Hungary; Molecular Animal Biotechnology Lab, Szent István University, Gödöllő, Hungary. Electronic address: Manuscript.Dinnyes@biotalentum.hu.

2017-11-13

Published in:

Stem cell research. - 2017

Induced Pluripotent Stem Cells

English Neural progenitor cells (NPCs) from human induced pluripotent stem cells (hiPSCs) are frequently induced using 3D culture methodologies however, it is unknown whether spheroid-based (3D) neural induction is actually superior to monolayer (2D) neural induction. Our aim was to compare the efficiency of 2D induction with 3D induction method in their ability to generate NPCs, and subsequently neurons and astrocytes. Neural differentiation was analysed at the protein level qualitatively by immunocytochemistry and quantitatively by flow cytometry for NPC (SOX1, PAX6, NESTIN), neuronal (MAP2, TUBB3), cortical layer (TBR1, CUX1) and glial markers (SOX9, GFAP, AQP4). Electron microscopy demonstrated that both methods resulted in morphologically similar neural rosettes. However, quantification of NPCs derived from 3D neural induction exhibited an increase in the number of PAX6/NESTIN double positive cells and the derived neurons exhibited longer neurites. In contrast, 2D neural induction resulted in more SOX1 positive cells. While 2D monolayer induction resulted in slightly less mature neurons, at an early stage of differentiation, the patch clamp analysis failed to reveal any significant differences between the electrophysiological properties between the two induction methods. In conclusion, 3D neural induction increases the yield of PAX6+/NESTIN+ cells and gives rise to neurons with longer neurites, which might be an advantage for the production of forebrain cortical neurons, highlighting the potential of 3D neural induction, independent of iPSCs' genetic background.

Language